scholarly journals Analysis of molecular weight albumin concentrate on various types of freshwater fish using SDS-page electrophoresis method

Author(s):  
Nurfaidah ◽  
Metusalach ◽  
Meta Mahendradatta ◽  
Sukarno
2019 ◽  
Vol 2 (1) ◽  
pp. 61
Author(s):  
Gunanti Mahasri, Ulia Fajriah, Sri Subekti

Abstract One of the diseases that often attack are fish is parasitic disease caused by Lernaea cyprinacea, called Lernaeosis. This ectoparasites could be detected on skin, gill, eyes, fin or inside of mouth and nostril of fishes. The aim of the study was to know the character is tic of L.cyprinacea protein based on molecular weight using SDS-PAGE. The results of this study were expected as Lernaeosis vaccine candidate. The experiment used SDS-PAGE with 12% separating gel, 3% stacking gel, voltage 110 volts 28 A and gel staining using coomassie blue. The result showed that, there were eight protein bands with molecular weight 82.3, 73.3, 66.6, 60.5, 54.9, 27.5, 23.1 and 19.8 kDa. Among them, the bands with molecular weight 27.5, 23.1 and 19.8 kDa were thicker and clearer than the others.


2019 ◽  
Vol 2 (1) ◽  
pp. 126
Author(s):  
Suardi Suardi ◽  
Ana Hidayanti Mukarromah ◽  
Stalis Norma Ethica

Fish is a potential source of animal protein, but has a weakness that is easy to rot. To avoid decay can be preserved by salting the fish. In this study wet salting was carried out to analyze the effect of salting on fish protein. The sample used was a type of gourami with 5 tails. 1 for the sample before salting and 4 for salting each salted at 10, 20, 30 and 40% b/v after that it was left to stand for 12 hours. The research method used was the Gel Electrophoresis method (SDS-PAGE) to determine molecular weight (MW), looking at the purity and damage of proteins in the sample. The samples of gouramy before salting showed 16 bands, 8 major bands and 8 minor ribbons. Samples of gouramy with a salt concentration of 10% b/v showed 16 bands, 7 major bands and 9 minor bands. Samples of gouramy with a salt concentration of 20% b/v showed 14 bands, 7 major bands and 7 minor bands. Samples of gouramy with a salt concentration of 30% b/v showed 10 bands, 3 major bands and 7 minor bands. Samples of gouramy with a salt concentration of 40% b/v showed 9 bands, 3 major bands and 6 minor bands. Thus it can be concluded that salting of fish can affect the gouramy protein, which is the higher the salt content added, the protein found in the fish will be denatured. The salting process of 10% b/v in gouramy meat is the most recommended salting process compared to the salting process of 20, 30 and 40% b/v.Ikan merupakan sumber protein hewani yang potensial, namun memiliki suatu kelemahan yaitu mudah membusuk. Untuk menghindari pembusukan dapat dilakukan pengawetan dengan penggaraman pada ikan. Pada penelitian ini dilakukan penggaraman basah untuk menganalisis pengaruh penggaraman terhadap protein ikan. Sampel yang digunakan adalah jenis ikan gurami sebanyak 5 ekor. 1 ekor untuk sampel sebelum penggaraman dan 4 ekor dilakukan penggaraman masing-masing digarami dengan kadar 10, 20, 30 dan 40% b/v setelah itu didiamkan selama 12 jam. Metode penelitian yang digunakan adalah metode Elektroforesis Gel (SDS- PAGE) untuk menentukan berat molekul (BM), melihat kemurnian dan kerusakan protein pada sampel. Pada sampel ikan gurami sebelum penggaraman menunjukkan 16 pita, 8 pita mayor dan 8 pita minor. Sampel ikan gurami dengan konsentrasi garam 10% b/v menujukkan 16 pita, 7 pita mayor dan 9 pita minor. Sampel ikan gurami dengan konsentrasi garam 20% b/v menunjukkan 14 pita, 7 pita mayor dan 7 pita minor. Sampel ikan gurami dengan konsentrasi garam 30% b/v menunjukkan 10 pita, 3 pita mayor dan 7 pita minor. Sampel ikan gurami dengan konsentrasi garam 40% b/v menunjukkan 9 pita, 3 pita mayor dan 6 pita minor. Dengan demikian dapat disimpulkan bahwa penggaraman pada ikan dapat berpengaruh terhadap protein ikan gurami yaitu makin tinggi kadar garam yang ditambahkan maka protein yang terdapat pada ikan akan terdenaturasi. Proses penggaraman 10% b/v pada daging ikan gurami merupakan proses penggaraman yang paling disarankan dibandingkan proses penggaraman 20,30 dan 40% b/v.


Author(s):  
Greffa J ◽  
Barrionuevo A ◽  
Vilcacundo E ◽  
Carrillo W

Objective: The aim of this study was to obtain kahai protein concentrate from Caryodendron orinocense karst cultivated in the region Amazonia of Ecuador and characterizes its gastric and duodenal hydrolysates using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) electrophoresis method and the reversed-phase ultra-high-performance liquid chromatography (RP-UHPLC) method.Methods: Kahai seeds (C. orinocense karst) were utilized to obtain kahai protein concentrate at pH 5.0 using the isoelectric precipitation method and then subject to gastric hydrolysis with pepsin enzyme (2000 U/mg of protein) at pH 1.2, pH 2.0, and pH 3.2 at 37°C for 2 h with agitation in simulated gastric fluids and then to duodenal hydrolysis with pancreatin (mix enzymes) at pH 7.0 at 37°C for 3 h with agitation in simulated intestinal fluid. Gastric and duodenal hydrolysates from kahai were characterized using the SDS-PAGE electrophoresis method and the RP-UHPLC chromatography method.Results: Proteins obtained from kahai (C. orinocense karst) were hydrolyzed with pepsin, only one protein with molecular weight of 100 kDa presented resistance to hydrolysis with pepsin at all pHs assayed. All proteins from kahai protein concentrate were totally hydrolyzed with pancreatin in in vitro conditions.Conclusion: This study suggests that kahai protein concentrates have a high grade of digestibility in vitro when using the gastroduodenal model of digestion. Kahai protein can be a good source of alternative vegetal proteins to be consumed by animals and humans.


2015 ◽  
Vol 77 (33) ◽  
Author(s):  
Anida Aminudin ◽  
Shamsul Muhammad ◽  
Ruhil Hayati Hamdan ◽  
Rumaizi Shaari ◽  
Mariam Firdhaus Mad Nordin ◽  
...  

Collagen was documented as a difficult and expensive protein to quantify due to its insoluble. It is particularly since solubility is a key functional property in a variety of applications such as healthcare and cosmetic products. The main aim of this study was to extract the collagens from the skins of commercial freshwater fish such as red tilapia (Oreochromis niloticus), catfish (Clarias gariepinus) and pangasius (Pangasius pangasius) together with characteristics defined for each type of collagen extraction. The extracted collagens were determined in their molecular weights by using SDS-PAGE and the structure of it was observed under SEM. The obtained molecular weight of all three commercial fish collagens is approximately >80kDa. The characteristics of each type of collagen were then defined by using appropriate analysis through this research.


2022 ◽  
Vol 2160 (1) ◽  
pp. 012009
Author(s):  
Tian Jin ◽  
Tao Wei ◽  
Zitong Zhang ◽  
Zehao Lei ◽  
Liquan Sun

Abstract There are two kinds of proteins in silk, sericin and silk fibroin. Polypeptide compounds from silk sericin and silk fibroin were prepared by hydrothermal method. The process of silk dissolution was investigated under different solid-liquid ratio, reaction temperature and reaction time. By controlling the operating parameters of hydrothermal method, the temperature, material ratio and time were further optimized, and the best experimental results were obtained, the expected decomposition of silks occurred when the ratio of silks to waters was selected as 1 to 10, at 140 degree in 30 min. The molecular weight of polypeptide was detected by SDS-PAGE electrophoresis and analysed by MALDI-TOF-MS. The results showed that the molecular weight of the polypeptide obtained from silks was about 6000-8000Da. After literature research, the polypeptide with such molecular weight could have better performance for some functional additives.


1999 ◽  
Vol 82 (11) ◽  
pp. 1428-1432 ◽  
Author(s):  
Cheryl Scott ◽  
Francesco Salerno ◽  
Elettra Lorenzano ◽  
Werner Müller-Esterl ◽  
Angelo Agostoni ◽  
...  

SummaryLittle is known about the regulation of high-molecular-weight-kininogen (HK) and low-molecular-weight-kininogen (LK) or the relationship of each to the degree of liver function impairment in patients with cirrhosis. In this study, we evaluated HK and LK quantitatively by a recently described particle concentration fluorescence immunoassay (PCFIA) and qualitatively by SDS PAGE and immunoblotting analyses in plasma from 33 patients with cirrhosis presenting various degrees of impairment of liver function. Thirty-three healthy subjects served as normal controls. Patients with cirrhosis had significantly lower plasma levels of HK (median 49 μg/ml [range 22-99 μg/ml]) and LK (58 μg/ml [15-100 μg/ml]) than normal subjects (HK 83 μg/ml [65-115 μg/ml]; LK 80 μg/ml [45-120 μg/ml]) (p < 0.0001). The plasma concentrations of HK and LK were directly related to plasma levels of cholinesterase (P < 0.0001) and albumin (P < 0.0001 and P < 0.001) and inversely to the Child-Pugh score (P < 0.0001) and to prothrombin time ratio (P < 0.0001) (reflecting the clinical and laboratory abnormalities in liver disease). Similar to normal individuals, in patients with cirrhosis, plasma HK and LK levels paralleled one another, suggesting that a coordinate regulation of those proteins persists in liver disease. SDS PAGE and immunoblotting analyses of kininogens in cirrhotic plasma showed a pattern similar to that observed in normal controls for LK (a single band at 66 kDa) with some lower molecular weight forms noted in cirrhotic plasma. A slight increase of cleavage of HK (a major band at 130 kDa and a faint but increased band at 107 kDa) was evident. The increased cleavage of HK was confirmed by the lower cleaved kininogen index (CKI), as compared to normal controls. These data suggest a defect in hepatic synthesis as well as increased destructive cleavage of both kininogens in plasma from patients with cirrhosis. The decrease of important regulatory proteins like kininogens may contribute to the imbalance in coagulation and fibrinolytic systems, which frequently occurs in cirrhotic patients.


1992 ◽  
Vol 68 (05) ◽  
pp. 534-538 ◽  
Author(s):  
Nobuhiko Yoshida ◽  
Shingi Imaoka ◽  
Hajime Hirata ◽  
Michio Matsuda ◽  
Shinji Asakura

SummaryCongenitally abnormal fibrinogen Osaka III with the replacement of γ Arg-275 by His was found in a 38-year-old female with no bleeding or thrombotic tendency. Release of fibrinopeptide(s) by thrombin or reptilase was normal, but her thrombin or reptilase time in the absence of calcium was markedly prolonged and the polymerization of preformed fibrin monomer which was prepared by the treatment of fibrinogen with thrombin or reptilase was also markedly defective. Propositus' fibrinogen had normal crosslinking abilities of α- and γ-chains. Analysis of fibrinogen chains on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the system of Laemmli only revealed the presence of abnormal γ-chain with an apparently higher molecular weight, the presence of which was more clearly detected with SDS-PAGE of fibrin monomer obtained by thrombin treatment. Purified fragment D1 of fibrinogen Osaka III also seemed to contain an apparently higher molecular weight fragment D1 γ remnant on Laemmli gels, which was digested faster than the normal control by plasmin in the presence of [ethy-lenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA).


1989 ◽  
Vol 61 (03) ◽  
pp. 437-441 ◽  
Author(s):  
Cindra Condra ◽  
Elka Nutt ◽  
Christopher J Petroski ◽  
Ellen Simpson ◽  
P A Friedman ◽  
...  

SummaryThe present work reports the discovery and charactenzation of an anticoagulant protein in the salivary gland of the giant bloodsucking leech, H. ghilianii, which is a specific and potent inhibitor of coagulation factor Xa. The inhibitor, purified to homogeneity, displayed subnanomolar inhibition of bovine factor Xa and had a molecular weight of approximately 15,000 as deduced by denaturing SDS-PAGE. The amino acid sequence of the first 43 residues of the H. ghilianii derived inhibitor displayed a striking homology to antistasin, the recently described subnanomolar inhibitor of factor Xa isolated from the Mexican leech, H. officinalis. Antisera prepared to antistasin cross-reacted with the H. ghilianii protein in Western Blot analysis. These data indicate that the giant Amazonian leech, H. ghilianii, and the smaller Mexican leech, H. officinalrs, have similar proteins which disrupt the normal hemostatic clotting mechanisms in their mammalian host’s blood.


Author(s):  
Tae Ito ◽  
Yuko Suzuki ◽  
Hideto Sano ◽  
Naoki Honkura ◽  
Francis J Castellino ◽  
...  

Background: Details of the molecular interaction between tissue type plasminogen activator (tPA) and plasminogen activator inhibitor type-1 (PAI-1) remain unknown. Methods and Results: Three distinct forms of high molecular weight complexes are demonstrated. Two of the forms were detected by mass spectrometry. The high molecular mass detected by MALDI-TOF MS spectrometry was 107,029 Da, which corresponds to the sum of molecular masses of the intact tPA (65,320 Da) and the intact PAI-1 (42,416 Da). The lower molecular mass was 104,367 Da and is proposed to lack the C-terminal bait peptide of PAI-1 (calculated mass, 3,804 Da) which was detected as a 3,808 Da fragment. When the complex was analyzed by SDS-PAGE, only a single band was observed. However, after treatment by SDS and Triton X-100, two distinct forms of the complex with different mobilities were shown by SDS-PAGE. The higher molecular weight band demonstrated specific tPA activity on fibrin autography, whereas the lower molecular weight band did not. Peptide sequence analysis of these two bands, however, unexpectedly revealed the existence of the C-terminal cleavage peptide in both bands and its amount was less in the upper band. In the upper band, the sequences corresponding to the regions at the interface between two molecules in its Michaelis intermediate were diminished. Thus, these two bands corresponded to distinct nonacyl-enzyme complexes, wherein only the upper band liberated free tPA under the conditions employed. Conclusion: These data suggest that under physiological conditions a fraction of the tPA-PAI-1 population exists as non-acylated-enzyme inhibitor complex.


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